Identification of potential toxicants in sediments from an industrialized area in Pohang, South Korea: Application of a cell viability assay of microalgae using flow cytometry

Potential toxicants in sediments collected from an industrialized bay of Korea were identified by use of effect-directed analysis (EDA). Three marine microalgal bioassays (Dunaliella tertiolecta, Isochrysis galbana, and Phaeodactylum tricornutum) with diverse endpoints were employed. Initial screening of raw organic extracts of sediments indicated large variations among locations and species in a traditional endpoint "inhibition of growth". After fractionation, inhibition of growths increased significantly, particularly in some fractions containing aromatics with log K_OW 5-6 (F2.6). While viabilities of cells were adversely affected in more fractions, including F2.6-F2.7 (log K_OW 5-7) and F3.5-F3.6 (log K_OW 4-6). Among the several endpoints of viability, esterase activity seemed to be more sensitive, followed by integrity of cell membranes, chlorophyll a, cell size, and intracellular complexity. Instrumental analyses indicated that toxicities to microalgae observed in F2.7 could not be fully explained by target PAHs. Full-scan screening analysis using GC-QTOFMS identified 58 compounds in F2.7 with matching scores ≥90%. Based on toxic potencies for these compounds predicted by ECOSAR, several causative agents, including 1-phenylpyrene, dibenz[a,c]anthracene, and picene were suggested. Overall, viability of microalgae provided sensitive and high-resolution toxicity screening of samples into integrative assessment of sediment.